Retention of LPG expression following infection of BMMs and BMDCs.

Abstract

(A) Anti-LPG fluorescence intensity on a per-parasite basis. Anti-LPG intensity of WT metacyclic-stage parasites as well as at various time points after infection of PEMs was measured as described in Methods. As a negative control, the anti-LPG intensity of L. major lpg1- (open circles) was measured 0.3 hr after infection of PEMs. The gray line shows the mean anti-LPG intensity of ‘LPG-negative’ WT parasites plus two standard deviations, and any parasite with anti-LPG values below that line would be considered “LPG-negative”. Black bars represent geometric mean of the data for each sample. Data shown is pooled from at least two independent experiments. ***, P (B) Representative images comparing the LPG-positivity of purified metacyclics and parasites within PEMs, DCs, and BMMs at 72 hours post-infection. Parasite nuclei are shown in green, LPG is shown in red. Scale bar represents 5 μm. (C) Percent of parasites within the three host cell types that are LPG+ as determined by the ‘qualitative’ assay. PEM data, black diamonds, BMM data, gray squares, DC data, open triangles. N > 200 parasites per time point/cell type. (D) Quantitation of LPG on the surface of parasites within PEMs, BMMs, and DCs at 24 and 72 hours post infection. For comparison, anti-LPG intensity data for metacyclic-stage parasites is also shown. The gray line shows the cut-off for LPG-positivity. Black bars represent geometric mean of the data for each sample. n ≥ 3 experiments. N.S, not significant, *, P P P < 0.0001 (ANOVA).</p